title: Quantification of microRNA expression with next-generation sequencing.
abstract: Rapid advancement of next-generation sequencing technologies has made it possible to study expression profiles of microRNAs (miRNAs) comprehensively and efficiently. Multiplexing miRNA libraries by barcoding can significantly reduce sequencing cost per sample without compromising library quality. This unit provides a step-by-step protocol for isolating miRNAs and constructing multiplexed miRNA libraries. Also described is a custom computational pipeline for analyzing the multiplexed miRNA library sequencing reads generated by Illumina-based technology.
authors.affiliation: Department of Genetics, Harvard Medical School, Boston, MA, USA.
authors.forename: Seda
authors.initials: S
authors.lastname: Eminaga
authors.forename: Danos C
authors.initials: DC
authors.lastname: Christodoulou
authors.forename: Francois
authors.initials: F
authors.lastname: Vigneault
authors.forename: George M
authors.initials: GM
authors.lastname: Church
authors.forename: J G
authors.initials: JG
authors.lastname: Seidman
mesh_terms: D019295:Computational Biology / Q000379:methods; D020869:Gene Expression Profiling / Q000379:methods*; D015723:Gene Library; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods*; D035683:MicroRNAs / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification
chemical_list: D035683:MicroRNAs